Detection of SS-A/Ro and SS-B/La Autoantibodies Using Immunoblotting Procedure and Characterization of the Antigens from 293 Cells and KB Cells

Specific and sensitive assay was performed to detect both antiSS-A/Ro and antiSS-B/La antibodies in sera of patients with collagen diseases including SLE, PSS, etc. The SS-A/Ro and SS-B/La antigens were prepared from human spleen (HSE) and cultured human cell line (KB cells), while rabbit thymus extract (RTE) was used as SS-B/La antigen marker. The antigens were partially purified by DEAE cellulose column chromatography. The SS-A/Ro antibody was shown to react mainly with 58KDa peptide by means of immunoblotting. Sera containing both the SS-A/Ro and SS-B/La antibody reacted with 40KDa peptide of RTE, and 58KDa, 42KDa and 40KDa peptides of HSE. We found that some of SS-A/Ro antisera could further react with 64KDa peptide in HSE. The 58KDa peptide is rich in a cytoplasmic fraction of KB cells, and the 40KDa peptide in the nucleoplasmic fraction. KB cells are not less good source of the antigens than human spleen. Extracts of 293 cells (human embryonic kidney cells expressing adenovirus-5 El gene) were prepared by the same method from KB cells, though immunoblotting patterns of both SS-A/Ro and SS-B/La antigens of 293 cell extracts are similar to those of KB cells, the relative content of SS-B/La antigens in 293 cell extracts are decreased

National trends in the outcomes of subarachnoid haemorrhage and the prognostic influence of stroke centre capability in Japan: retrospective cohort study

Objectives To examine the national, 6-year trends in in-hospital clinical outcomes of patients with subarachnoid haemorrhage (SAH) who underwent clipping or coiling and the prognostic influence of temporal trends in the Comprehensive Stroke Center (CSC) capabilities on patient outcomes in Japan.Design Retrospective study.Setting Six hundred and thirty-one primary care institutions in Japan.Participants Forty-five thousand and eleven patients with SAH who were urgently hospitalised, identified using the J-ASPECT Diagnosis Procedure Combination database.Primary and secondary outcome measures Annual number of patients with SAH who remained untreated, or who received clipping or coiling, in-hospital mortality and poor functional outcomes (modified Rankin Scale: 3–6) at discharge. Each CSC was assessed using a validated scoring system (CSC score: 1–25 points).Results In the overall cohort, in-hospital mortality decreased (year for trend, OR (95% CI): 0.97 (0.96 to 0.99)), while the proportion of poor functional outcomes remained unchanged (1.00 (0.98 to 1.02)). The proportion of patients who underwent clipping gradually decreased from 46.6% to 38.5%, while that of those who received coiling and those left untreated gradually increased from 16.9% to 22.6% and 35.4% to 38%, respectively. In-hospital mortality of coiled (0.94 (0.89 to 0.98)) and untreated (0.93 (0.90 to 0.96)) patients decreased, whereas that of clipped patients remained stable. CSC score improvement was associated with increased use of coiling (per 1-point increase, 1.14 (1.08 to 1.20)) but not with short-term patient outcomes regardless of treatment modality.Conclusions The 6-year trends indicated lower in-hospital mortality for patients with SAH (attributable to better outcomes), increased use of coiling and multidisciplinary care for untreated patients. Further increasing CSC capabilities may improve overall outcomes, mainly by increasing the use of coiling. Additional studies are necessary to determine the effect of confounders such as aneurysm complexity on outcomes of clipped patients in the modern endovascular era